Search results for: combined aquatic and cellular toxicity

Authors: Najmeh Hoseini

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Strokerecovery happens through neuroplasticity, which is highly influenced by the environment, including neuro-rehabilitation. Transcranial direct current stimulation (tDCS) may enhance recovery by modulating neuroplasticity. With tDCS, weak direct currents are applied noninvasively to modify excitability in the cortical areas under its electrodes. Combined with functional activities, this may facilitate motor recovery in neurologic disorders such as stroke. The purpose of this case study was to examine the effect of tDCS combined with 30 minutes of traditional physical therapy (PT)on arm function following a stroke. A 29-year-old male with chronic stroke involving the left middle cerebral artery territory went through the treatment protocol. Design The design included 5 weeks of treatment: 1 week of traditional PT, 2 weeks of sham tDCS combined with traditional PT, and 2 weeks of tDCS combined with traditional PT. PT included functional electrical stimulation (FES) of wrist extensors followed by task-specific functional training. Dual hemispheric tDCS with 1 mA intensity was applied on the sensorimotor cortices for the first 20 min of the treatment combined with FES. Assessments before and after each treatment block included Modified Ashworth Scale, ChedokeMcmaster Arm and Hand inventory, Action Research Arm Test (ARAT), and the Box and Blocks Test. Results showed reduced spasticity in elbow and wrist flexors only after tDCS combination weeks (+1 to 0). The patient demonstrated clinically meaningful improvements in gross motor and fine motor control over the duration of the study; however, components of the ARAT that require fine motor control improved the greatest during the experimental block. Average time improvement compared to baseline was26.29 s for tDCS combination weeks, 18.48 s for sham tDCS, and 6.83 for PT standard of care weeks. Combining dual hemispheric tDCS with the standard of care PT demonstrated improvements in hand dexterity greater than PT alone in this patient case.

Keywords: tDCS, stroke, case study, physical therapy

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Authors: Nomthandazo Precious Sibiya, Thembisile Patience Mahlangu, Sudesh Rathilal

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Heavy metal removal is critical in the wastewater treatment process due to its numerous harmful effects on human and aquatic life. There are several chemical and physical techniques for removing heavy metals from wastewater, including ion exchange, reverse osmosis, adsorption, electrodialysis, and ultrafiltration. However, adsorption technology has captivated researchers for years due to its low cost, high efficiency, and compatible with the environment. In this study, the adsorption effectiveness of three modified agro-waste materials was explored for the removal of lead from synthetic wastewater: banana peels (BP), orange peels (OP), and sugarcane bagasse (SB). The magnetite (Fe₃O₄) is incorporated with BP, OP, and SB at a ratio of 1:1 to create magnetic biosorbents. Characterization of biosorbents was carried out using and scanning electron microscopy (SEM) combined with energy-dispersive X-ray (EDX) to investigate surface morphology and elemental compositions, respectively. A series of batch experiments were carried out to investigate the effects of adsorbent mass, agitation time, and initial pH concentration on adsorption behaviour, as well as adsorption isotherms and kinetics. The removal efficiency of lead by the modified agro-waste materials proved to be superior to that of non-modified agro-waste materials. The proof of concept was achieved, and agro-waste materials can be paired with adsorption technology to effectively remove lead from aqueous media. The use of agricultural waste as biosorbents will aid in waste reduction and management.

Keywords: adsorption, isotherms, kinetics, agro waste, nanoparticles, batch

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Authors: Alemu Tadesse Feroche

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In this study, some 2, 3 distributed quinazoline -4 (3H) - one derivative were synthesized using a three-step synthetic route. They were obtained in a good yield (59.5-85%) by applying different chemical reactions like cyclization and condensation reactions. The chemical structure of the final compounds was also verified by spectroscopic methods (IR, ¹HNMR) and elemental microanalysis. The in vivo antimalarial activity of these compounds on P. berghei infected mice was found to be moderate to high at an oral dose of 0.04846 mmol/kg /day. This is equal to 25 mg/kg of chloroquine phosphate, which causes 100% inhibition of the parasite. It is worth mentioning that most active compounds (E) -3 Phenyl -2- [2- (pyridine -4- yl) vinyl] -4 (3H) -quinazolinone IVa (64.02%, (E)-2-[2-(4 - Hydroxy-3 - methoxystyryl) - vinyl) -3 - phenyl -4 (3H ) - quinazolinone IVc (77.25%) and (E)-2 –[2 –(Pyridin -4-yl) –vinyl] -3 phenenylamine -4(3H) quinazolinone IVe (73.54%) showed a dose-dependent increase in present suppression in antimalarial activities. Furthermore, the synthesized compounds were screened for their in vitro antileishmanial activity against L. aethiopica isolate (CL/039/09). All tested compounds (IVa (0.03766 ug/ml), IVb (0.00538 ug/ml, IVc (0.00412 ug/ml, IVd (0.00110 ug/ml), IVe (0.03017 ug/ml) and IVf (0.03894 ug/ml)) showed excellent potency that is much better than amphotericin B (IC50 = 0,04359 ug/ml). The results of acute toxicity indicated that all test compounds (IVa –IVf) proved to be nontoxic and well tolerated by the experimental animals up to 300 mg/kg in oral and 140 mg/kg in parental studies.

Keywords: 4(3H)-quinazolinone, in vivo antimalarial activity, in vitro antileishmanial activity, acute toxicity

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Authors: Komal Saini, S. K. Sahoo, B. S. Bajwa

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The Laser Fluorimetery Technique has been used for the microanalysis of uranium content in water samples collected from different sources like the hand pumps, tube wells in the drinking water samples of SW & NE Punjab, India. The geographic location of the study region in NE Punjab is between latitude 31.21º- 32.05º N and longitude 75.60º-76.14º E and for SW Punjab is between latitude 29.66º-30.48º N and longitude 74.69º-75.54º E. The purpose of this study was mainly to investigate the uranium concentration levels of ground water being used for drinking purposes and to determine its health effects, if any, to the local population of these regions. In the present study 131 samples of drinking water collected from different villages of SW and 95 samples from NE, Punjab state, India have been analyzed for chemical and radiological toxicity. In the present investigation, uranium content in water samples of SW Punjab ranges from 0.13 to 908 μgL−1 with an average of 82.1 μgL−1 whereas in samples collected from NE- Punjab, it ranges from 0 to 28.2 μgL−1 with an average of 4.84 μgL−1. Thus, revealing that in the SW- Punjab 54 % of drinking water samples have uranium concentration higher than international recommended limit of 30 µgl-1 (WHO, 2011) while 35 % of samples exceeds the threshold of 60 µgl-1 recommended by our national regulatory authority of Atomic Energy Regulatory Board (AERB), Department of Atomic Energy, India, 2004. On the other hand in the NE-Punjab region, none of the observed water sample has uranium content above the national/international recommendations. The observed radiological risk in terms of excess cancer risk ranges from 3.64x10-7 to 2.54x10-3 for SW-Punjab, whereas for NE region it ranges from 0 to 7.89x10-5. The chemical toxic effect in terms of Life-time average Daily Dose (LDD) and Hazard Quotient (HQ) have also been calculated. The LDD for SW-Punjab varies from 0.0098 to 68.46 with an average of 6.18 µg/ kg/day whereas for NE region it varies from 0 to 2.13 with average 0.365 µg/ kg/day, thus indicating presence of chemical toxicity in SW Punjab as 35% of the observed samples in the SW Punjab are above the recommendation limit of 4.53 µg/ kg/day given by AERB for 60 µgl-1 of uranium. Maximum & Minimum values for hazard quotient for SW Punjab is 0.002 & 15.11 with average 1.36 which is considerably high as compared to safe limit i.e. 1. But for NE Punjab HQ varies from 0 to 0.47. The possible sources of high uranium observed in the SW- Punjab will also be discussed.

Keywords: uranium, groundwater, radiological and chemical toxicity, Punjab, India

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Authors: Md. Imran H. Khan, T. Farrell, M. A. Karim

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Highly porous and hygroscopic characteristics of pear make it complex to understand the cellular level water distribution. In pear tissue, water is mainly distributed in three different spaces namely, intercellular water, intracellular water, and cell wall water. Understanding of these three types of water in pear tissue is crucial for predicting actual heat and mass transfer during drying. Therefore, the aim of the present study was to investigate the proportion of intercellular water, intracellular water, and cell wall water inside the pear tissue. During this study, Green Anjou Pear was taken for the investigation. The experiment was performed using 1H-NMR- T2 relaxometry. Various types of water component were calculated by using multi-component fits of the T2 relaxation curves. The experimental result showed that in pear tissue 78-82% water exist in intracellular space; 12-16% water in intercellular space and only 2-4% water exist in the cell wall space. The investigated results quantify different types of water in plant-based food tissue. The highest proportion of water exists in intracellular spaces. It was also investigated that the physical properties of pear and the proportion of the different types of water has a strong relationship. Cell wall water depends on the proportion of solid in the sample tissue whereas free water depends on the porosity of the material.

Keywords: intracellular water, intercellular water, cell wall water, physical property, pear

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Authors: Farahnaz Motamedi Sedeh, Homayoon Mahravani, Parvin Shawrang, Mehdi Behgar

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Most countries use inactivated binary ethylenimine (BEI) vaccines to control and prevent Foot-and-Mouth Disease (FMD). However, this vaccine induces a short-term humoral immune response in animals. This study investigated the cellular and humoral immune responses in homologous and prime-boost (PB) groups in the BALB/c mouse model. FMDV strain O/IRN/1/2007 was propagated in the BHK-21 cell line and inactivated by three methods, including a chemical with BEI to produce a conventional vaccine (CV), a gamma irradiation vaccine (GIV), and an electron irradiated vaccine (EIV). Three vaccines were formulated with the adjuvant aluminum hydroxide gel. In addition, a DNA vaccine was prepared by amplifying the virus VP1 gene pcDNA3.1 plasmid. In addition, the plasmid encoding the granulocyte-macrophage colony-stimulating factor gene (GM-CSF) was used as a molecular adjuvant. Eleven groups of five mice each were selected, and the vaccines were administered as homologous and heterologous strategy prime-boost (PB) in three doses two weeks apart. After the evaluation of neutralizing antibodies, interleukin (IL)-2, IL-4, IL-10, interferon-gamma (INF-γ), and MTT assays were compared in the different groups. The pcDNA3.1+VP1 cassette was prepared and confirmed as a DNA vaccine. The virus was inactivated by gamma rays and electron beams at 50 and 55 kGy as GIV and EIV, respectively. Splenic lymphocyte proliferation in the inactivated vaccinated homologous groups was significantly lower (P≤0.05) compared with the heterologous prime-boosts (PB1, PB2, PB3) and DNA + GM-CSF groups (P≤0.05). The highest SNT titer was observed in the inactivated vaccine groups. IFN-γ and IL-2 were higher in the vaccinated groups. It was found that although there was a protective humoral immune response in the groups with inactivated vaccine, there was adequate cellular immunity in the group with the DNA vaccine. However, the strongest cellular and humoral immunity was observed in the PB groups. The primary injection was accompanied by DNA vaccine + GM-CSF and boosted injection with GIV or CV.

Keywords: foot and mouth disease, irradiated vaccine, immune responses, DNA vaccine, prime boost strategy

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Authors: Safee Chaudhary, Mahnoor Naseer Gondal, Hira Anees Awan, Abdul Rehman, Ammar Arif, Risham Hussain, Huma Khawar, Zainab Arshad, Muhammad Faizyab Ali Chaudhary, Waleed Ahmed, Muhammad Umer Sultan, Bibi Amina, Salaar Khan, Muhammad Moaz Ahmad, Osama Shiraz Shah, Hadia Hameed, Muhammad Farooq Ahmad Butt, Muhammad Ahmad, Sameer Ahmed, Fayyaz Ahmed, Omer Ishaq, Waqar Nabi, Wim Vanderbauwhede, Bilal Wajid, Huma Shehwana, Muhammad Tariq, Amir Faisal

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Cancer is a manifestation of multifactorial deregulations in biomolecular pathways. These deregulations arise from the complex multi-scale interplay between cellular and extracellular factors. Such multifactorial aberrations at gene, protein, and extracellular scales need to be investigated systematically towards decoding the underlying mechanisms and orchestrating therapeutic interventions for patient treatment. In this work, we propose ‘TISON’, a next-generation web-based multiscale modeling platform for clinical systems oncology. TISON’s unique modeling abstraction allows a seamless coupling of information from biomolecular networks, cell decision circuits, extra-cellular environments, and tissue geometries. The platform can undertake multiscale sensitivity analysis towards in silico biomarker identification and drug evaluation on cellular phenotypes in user-defined tissue geometries. Furthermore, integration of cancer expression databases such as The Cancer Genome Atlas (TCGA) and Human Proteome Atlas (HPA) facilitates in the development of personalized therapeutics. TISON is the next-evolution of multiscale cancer modeling and simulation platforms and provides a ‘zero-code’ model development, simulation, and analysis environment for application in clinical settings.

Keywords: systems oncology, cancer systems biology, cancer therapeutics, personalized therapeutics, cancer modelling

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Authors: Priyanka Chowdhury, Asitikantha Sarma, Utpal Ghosh

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Hadron therapy using high Linear Energy Transfer (LET) ion beam is producing promising clinical results worldwide. The major advantages are its ability to kill radio-resistant tumor and its anti-metastatic activity. Poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors have been widely used as radiosensitizer, but its role in metastasis is unknown. The purpose of our study was to investigate the effect of PARP-1 depletion in combination with either Carbon Ion Beam (CIB) or gamma irradiation on metastatic potential of cultured cancerous cells. A549 cells were irradiated with CIB (0-4Gy) or gamma (0, 2, 4, 6 and 10 Gy) with and without PARP-1 inhibition. The metastatic potential of the cells was determined by cell migratory assay, expression, and activity of MMP-2 and MMP-9, expression of Cadherin, Fibronectin, and Vimentin. CIB exposure reduced migratory property and activity of MMP-2 and MMP-9 significantly. CIB with PARP-1 inhibition reduced cell migration and Matrix Metalloproteinase (MMPs) activity in a synergistic manner. Expression of MMPs was also down-regulated in CIB and combined treatment. On the contrary, MMP- 2 and MMP-9 activity was significantly increased in gamma irradiated cells but decreased upon combined treatment of gamma and PARP-1 inhibitor. MMPs expression and migration was reduced when gamma irradiation was combined with PARP-1 inhibition. Thus, our study clearly demonstrates that PARP-1 inhibition in combination with either high or low LET can significantly suppress metastatic potential in cancer cells and thereby can be a promising tool in controlling metastatic cancers.

Keywords: high LET, low LET, matrix metalloproteinase (MMP), PARP-1

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Authors: Surbhi Surbhi, Andrea Erni, Gunter Meister, Harold Cremer, Christophe Beclin

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MicroRNAs (miRNAs) are short 20-23 nucleotide long non-coding RNAs; there are 2605 miRNA in humans and 1936 miRNA in mouse in total (miRBase). The nervous system expresses the most abundant miRNA and most diverse. MiRNAs play a role in many steps during neurogenesis, like cell proliferation, differentiation, neural patterning, axon pathfinding, etc. Moreover, in vitro studies suggested a role in the regulation of local translation at the synapse, thus controlling neuronal plasticity. However, due to the specific structure of miRNA molecules, an in-vivo confirmation of the general role of miRNAs in the control of neuronal plasticity is still pending. For example, their small size and their high level of sequence homology make difficult the analysis of their cellular and sub-cellular localization in-vivo by in-situ hybridization. Moreover, it was found that only 40% of the expressed miRNA molecules in a cell are included in RNA-Induced Silencing Complexes (RISC) and, therefore, involved in inhibitory interactions while the rest is silent. Definitively, the development of new tools is needed to have a better understanding of the cellular function of miRNAs, in particular their role in neuronal plasticity. Here we describe a new technique called in-vivo AGO-APP designed to investigate miRNA expression and function in-vivo. This technique is based on the expression of a small peptide derived from the human RISC-complex protein TNRC6B, called T6B, which binds all known Argonaute (Ago) proteins with high affinity allowing the efficient immunoprecipitation of AGO-bound miRNAs. We have generated two transgenic mouse lines conditionally expressing T6B either ubiquitously in the cell or targeted at the synapse. A comparison of the repertoire of miRNAs immuno-precipitated from mature neurons of both mouse lines will provide us with a list of miRNAs showing a specific activity at the synapse. The physiological role of these miRNAs will be subsequently addressed through gain and loss of function experiments.

Keywords: RNA-induced silencing complexes, TNRC6B, miRNA, argonaute, synapse, neuronal plasticity, neurogenesis

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Authors: Vikash Yadav, Ashish Arora

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Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

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Authors: Dong-Gi Lee, Suyeon Cha, Yong-Sun Bahn

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Sterol lipid is essential for cell membrane structure in eukaryotic cells. In mammalian cells, sterol regulatory element binding proteins (SREBPs) act as principal regulators of cellular cholesterol which is essential for proper cell membrane fluidity and structure. SREBP and sterol regulation are related to levels of cellular oxygen because it is a major substrate for sterol synthesis. Upon cellular sterol and oxygen levels are depleted, SREBP is translocated to the Golgi where it undergoes proteolytic cleavage of N terminus, then it travels to the nucleus to play a role as transcription factor. In yeast cells, synthesis of ergosterol is also highly oxygen consumptive, and Sre1 is a transcription factor known to play a central role in adaptation to growth under low oxygen condition and sterol homeostasis in Cryptococcus neoformans. In this study, we observed phenotypes in other strains of Cryptococcus species by constructing hob1Δ and sre1Δ mutants to confirm whether the functions of both genes are conserved in most serotypes. As a result, hob1Δ showed no noticeable phenotype under treatment of antifungal drugs and most environmental stresses in R265 (C. gattii) and XL280 (C. neoformans), suggesting that Hob1 is related to sterol regulation only in H99 (serotype A). On the other hand, the function of Sre1 was found to be conserved in most serotypes. Furthermore, mating experiment of hob1Δ or sre1Δ showed dramatic defects in serotype A (H99) and D (XL280). It revealed that Hob1 and Sre1 related to mating ability in Cryptococcus species, especially cell fusion efficiency. In conclusion, HOB1 and SRE1 play crucial role in regulating sterol-homeostasis and differentiation in C. neoformans, moreover, Hob1 is specific gene in Cryptococcus neoformans. It suggests that Hob1 is considered as potent factor-targeted new safety antifungal drug.

Keywords: cryptococcus neoformans, Hob1, Sre1, sterol regulatory element binding proteins

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Authors: Bouafia-Chergui Souâd, Chabani Malika, Bensmaili Aicha

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Water treatment and especially, medicament pollutants are nowadays important problems. Degradation of oxytetracycline was carried out using combined process of low-frequency ultrasound (US), ultraviolet irradiation and a catalyst. The effectiveness of the coupled processes has been evaluated by studying the effects of various operating parameters including initial OTC concentration, solution pH and catalyst mass. For the photolysis process, the monochromatic ultraviolet light wavelength utilized was 365 nm. The sonolysis experiments were performed with ultrasound at a frequency of 40 kHz. The heterogeneous photocatalysis was studied in the presence of TiO2. The processes were employed individually, and simultaneously to examine the details of the processes and to investigate the contribution of each process. Low UV intensity (12W), low pH and high mass of TiO2 conditions enhanced the sono-photocatalytic degradation of OTC. The results showed that the individual contribution sonochemical and photochemical reactions are very low, however, their coupling increases the degradation rate of 8 times compared to photolysis and 2 times compared to sonolysis. There is a synergistic effect between the two modes of radiation, UV and U.S. leading to 82.04% degradation yield. An application of these combined processes on the treatment of a real pharmaceutical wastewater was examined.

Keywords: sonolysis, photocatalysis, combined process, antibiotic

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Authors: P. H. Mfengwana, S. S. Mashele, L. Verschaeve, R. Anthonissen, I. T. Manduna

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Gunnera perpensa is traditionally used mostly by women for the treatment of different gynaecological related conditions due to its proven uterine contractility effects. The uses of this plant include menstrual pain relief, treatment of infertility and promotion of easy labour. However, even though this plant species has been reported to possess numerous medicinal properties, to author’s best knowledge, its safety has not been investigated. Thus, this study was aimed at investigating the genotoxicity and antigenotoxicity of Gunnera perpensa aqueous, methanol and dichloromethane extracts. The in vitro toxicity of the plant extracts was assessed with the neutral red uptake (NRU) test. Genotoxic and antigenotoxic properties of Gunnera perpensa were investigated using high-throughput assays: bacterial Vitotox test and the alkaline comet assay with and without S9 activation on human C3A cells. Ethyl Methanesulfonate (EMS) and 4-nitroquinoline-oxide (4-NQO) were used as positive controls, respectively. All extracts showed toxicity in a dose-dependent manner; however, that does not mean they were all genotoxic. Methanol extract did show genotoxicity with S9 (metabolism) only at the highest concentration of 500 µg/ml due to increased DNA damage observed, however, no genotoxicity was observed from other concentrations. Therefore, the results show that Gunnera perpensa extracts are genotoxic and not safe for human use.

Keywords: antigenotoxicity, comet test, genotoxicity, Gunnera perpensa, vitotox assay

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Authors: Maryam Mohammad Hadi, Heather Nesbitt, Hamzah Masood, Hashim Ahmed, Mark Emberton, John Callan, Alexander MacRobert, Anthony McHale, Nikolitsa Nomikou

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Sonodynamic therapy (SDT) utilises ultrasound in combination with sensitizers, such as porphyrins, for the production of cytotoxic reactive oxygen species (ROS) and the confined ablation of tumours. Ultrasound can be applied locally, and the acoustic waves, at frequencies between 0.5-2 MHz, are transmitted efficiently through tissue. SDT does not require highly toxic agents, and the cytotoxic effect only occurs upon ultrasound exposure at the site of the lesion. Therefore, this approach is not associated with adverse side effects. Further highlighting the benefits of SDT, no cancer cell population has shown resistance to therapy-triggered ROS production or their cytotoxic effects. This is particularly important, given the as yet unresolved issues of radiation and chemo-resistance, to the authors’ best knowledge. Another potential future benefit of this approach – considering its non-thermal mechanism of action – is its possible role as an adjuvant to immunotherapy. Substantial pre-clinical studies have demonstrated the efficacy and targeting capability of this therapeutic approach. However, SDT has yet to be fully characterised and appropriately exploited for the treatment of cancer. In this study, a formulation based on multistimulus-responsive sensitizer-containing nanoparticles that can accumulate in advanced prostate tumours and increase the therapeutic efficacy of SDT has been developed. The formulation is based on a polyglutamate-tyrosine (PGATyr) co-polymer carrying hematoporphyrin. The efficacy of SDT in this study was demonstrated using prostate cancer as the translational exemplar. The formulation was designed to respond to the microenvironment of advanced prostate tumours, such as the overexpression of the proteolytic enzymes, cathepsin-B and prostate-specific membrane antigen (PSMA), that can degrade the nanoparticles, reduce their size, improving both diffusions throughout the tumour mass and cellular uptake. The therapeutic modality was initially tested in vitro using LNCaP and PC3 cells as target cell lines. The SDT efficacy was also examined in vivo, using male SCID mice bearing LNCaP subcutaneous tumours. We have demonstrated that the PGATyr co-polymer is digested by cathepsin B and that digestion of the formulation by cathepsin-B, at tumour-mimicking conditions (acidic pH), leads to decreased nanoparticle size and subsequent increased cellular uptake. Sonodynamic treatment, at both normoxic and hypoxic conditions, demonstrated ultrasound-induced cytotoxic effects only for the nanoparticle-treated prostate cancer cells, while the toxicity of the formulation in the absence of ultrasound was minimal. Our in vivo studies in immunodeficient mice, using the hematoporphyrin-containing PGATyr nanoparticles for SDT, showed a 50% decrease in LNCaP tumour volumes within 24h, following IV administration of a single dose. No adverse effects were recorded, and body weight was stable. The results described in this study clearly demonstrate the promise of SDT to revolutionize cancer treatment. It emphasizes the potential of this therapeutic modality as a fist line treatment or in combination treatment for the elimination or downstaging of difficult to treat cancers, such as prostate, pancreatic, and advanced colorectal cancer.

Keywords: sonodynamic therapy, nanoparticles, tumour ablation, ultrasound

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Authors: Medina O. Kadiri, John E. Gabriel

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Heavy metals are metallic elements with a relatively high density, at least five times greater compared to water. The sources of heavy metal pollution in the environment include industrial, medical, agricultural, pharmaceutical, domestic effluents, and atmospheric sources, mining, foundries, smelting, and any heavy metal-based operation. Although some heavy metals in trace quantities are required for biological metabolism, their higher concentrations elicit toxicities. Others are distinctly toxic and are of no biological functions. Microalgae are the primary producers of aquatic ecosystems and, therefore, the foundation of the aquatic food chain. A study investigating the effects of copper and zinc on the two chlorophytes-Chlorella vulgaris and Dictyosphaerium pulchellum was done in the laboratory, under different concentrations of 0mg/l, 2mg/l, 4mg/l, 6mg/l, 8mg/l, 10mg/l, and 20mg/l. The growth of the test microalgae was determined every two days for 14 days. The results showed that the effects of the test heavy metals were concentration-dependent. From the two microalgae species tested, Chlorella vulgaris showed appreciable growth up to 8mg/l concentration of zinc. Dictyoshphaerium pulchellum had only minimal growth at different copper concentrations except for 2mg/l, which seemed to have relatively higher growth. The growth of the control was remarkably higher than in other concentrations. Generally, the growth of both test algae was consistently inhibited by heavy metals. Comparatively, copper generally inhibited the growth of both algae than zinc. Chlorella vulgaris can be used for bioremediation of high concentrations of zinc. The potential of many microalgae in heavy metal bioremediation can be explored.

Keywords: heavy metals, green algae, microalgae, pollution

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Authors: Aliro Villacorta, Laura Rubio, Mohamed Alaraby, Montserrat López Mesas, Victor Fuentes-Cebrian, Oscar H. Moriones, Ricard Marcos, Alba Hernández.

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Micro and nano plastics (MNPLs) are emergent environmental pollutants requiring urgent information on their potential risks to human health. One of the problems associated with the evaluation of their undesirable effects is the lack of real samples matching those resulting from the environmental degradation of plastic wastes. To such end, we propose an easy method to obtain polyethylene terephthalate nano plastics from water plastic bottles (PET-NPLs) but, in principle, applicable to any other plastic goods sources. An extensive characterization indicates that the proposed process produces uniform samples of PET-NPLs of around 100 nm, as determined by using a multi-angle and dynamic light scattering methodology. An important point to be highlighted is that to avoid the metal contamination resulting from methods using metal blades/burrs for milling, trituration, or sanding, we propose to use diamond burrs to produce metal-free samples. To visualize the toxicological profile of the produced PET-NPLs, we have evaluated their ability to be internalized by cells, their cytotoxicity, and their ability to induce oxidative stress and induce DNA damage. In this preliminary approach, we have detected their cellular uptake, but without the induction of significant biological effects. Thus, no relevant increases in toxicity, reactive oxygen species (ROS) induction, or DNA damage -as detected with the comet assay- have been observed. The use of real samples, as produced in this study, will generate relevant data in the discussion about the potential health risks associated with MNPLs exposures.

Keywords: nanoplastics, polyethylene terephthalate, physicochemical characterization, cell uptake, cytotoxicity

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Authors: A. Somaya Madkour, B. Abeer sheta, C. Fatma Badr El Dine, D. Yasser Elwakeel, E. Nermine AbdAllah

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Criminal offenders have a fundamental goal not to leave any traces at the crime scene. Some may suppose that items recovered underwater will have no forensic value, therefore, they try to destroy the traces by throwing items in water. These traces are subjected to the destructive environmental effects. This can represent a challenge for Forensic experts investigating finger marks. Accordingly, the present study was conducted to determine the optimal method for latent fingerprints development on non-porous surfaces submerged in aquatic environments at different time interval. The two factors analyzed in this study were the nature of aquatic environment and length of submerged time. In addition, the quality of developed finger marks depending on the used method was also assessed. Therefore, latent fingerprints were deposited on metallic, plastic and glass objects and submerged in fresh or sea water for one, two, and ten days. After recovery, the items were subjected to cyanoacrylate fuming, black powder and small particle reagent processing and the prints were examined. Each print was evaluated according to fingerprint quality assessment scale. The present study demonstrated that the duration of submersion affects the quality of finger marks; the longer the duration, the worse the quality.The best results of visualization were achieved using cyanoacrylate either in fresh or sea water. This study has also revealed that the exposure to sea water had more destructive influence on the quality of detected finger marks.

Keywords: fingerprints, fresh water, sea, non-porous

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Authors: Adam M. Pitz, Gillian L. Phillipson, Jayant E. Khanolkar, Andrew M. Middleton

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New emerging evidence suggests that the nose is the predominant route for entry of the SARS-CoV-2 virus into the host. A virucidal suspension test (conforming in principle to the European Standard EN14476) was conducted to determine whether a commercial liquid gel intranasal spray containing 1% of the mucoadhesive hydroxypropyl methylcellulose (HPMC) could inhibit the cellular infectivity of the SARS-CoV-2 coronavirus. Virus was added to the test product samples and to controls in a 1:8 ratio and mixed with one part bovine serum albumin as an interfering substance. The test samples were pre-equilibrated to 34 ± 2°C (representing the temperature of the nasopharynx) with the temperature maintained at 34 ± 2°C for virus contact times of 1, 5 and 10 minutes. Neutralized aliquots were inoculated onto host cells (Vero E6 cells, ATCC CRL-1586). The host cells were then incubated at 36 ± 2°C for a period of 7 days. The residual infectious virus in both test and controls was detected by viral-induced cytopathic effect. The 50% tissue culture infective dose per mL (TCID50/mL) was determined using the Spearman-Karber method with results reported as the reduction of the virus titer due to treatment with test product, expressed as log10. The controls confirmed the validity of the results with no cytotoxicity or viral interference observed in the neutralized test product samples. The HPMC formulation reduced SARS-CoV-2 titer, expressed as log10TCID50, by 2.30 ( ± 0.17), 2.60 ( ± 0.19), and 3.88 ( ± 0.19) with the respective contact times of 1, 5 and 10 minutes. The results demonstrate that this 1% HPMC gel formulation can reduce the cellular infectivity of the SARS-CoV-2 virus with an increasing viral inhibition observed with increasing exposure time. This 1% HMPC gel is well tolerated and can reside, when delivered via nasal spray, for up to one hour in the nasal cavity. We conclude that this intranasal gel spray with 1% HPMC repeat-dosed every few hours may offer an effective preventive or early intervention solution to limit the transmission and impact of the SARS-CoV-2 coronavirus.

Keywords: hydroxypropyl methylcellulose, mucoadhesive nasal spray, respiratory viruses, SARS-CoV-2

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Authors: K. Verma, v. S. Moholkar

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In developed countries, water pollution caused by industrial discharge has emerged as a significant environmental concern over the past decades. However, despite ongoing efforts, a fully effective and sustainable remediation strategy has yet to be identified. This paper describes how enzymatic and sonochemical treatments have demonstrated great promise in degrading bio-refractory pollutants. Mainly, a compelling area of interest lies in the combined technique of sono-enzymatic treatment, which has exhibited a synergistic enhancement effect surpassing that of the individual techniques. This study employed the covalent attachment method to immobilize Laccase from Trametes versicolor onto amino-functionalized magnetic Fe₃O₄ nanoparticles. To comprehensively characterize the synthesized free nanoparticles and the laccase-immobilized nanoparticles, various techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), vibrating sample magnetometer (VSM), and surface area through Brunauer-Emmett-Teller (BET) were employed. The size of immobilized Fe₃O₄@Laccase was found to be 60 nm, and the maximum loading of laccase was found to be 24 mg/g of nanoparticle. An investigation was conducted to study the effect of various process parameters, such as immobilized Fe₃O₄ Laccase dose, temperature, and pH, on the % Chemical oxygen demand (COD) removal as a response. The statistical design pinpointed the optimum conditions (immobilized Fe₃O₄ Laccase dose = 1.46 g/L, pH = 4.5, and temperature = 66 oC), resulting in a remarkable 65.58% COD removal within 60 minutes. An even more significant improvement (90.31% COD removal) was achieved with ultrasound-assisted enzymatic reaction utilizing a 10% duty cycle. The investigation of various kinetic models for free and immobilized laccase, such as the Haldane, Yano, and Koga, and Michaelis-Menten, showed that ultrasound application impacted the kinetic parameters Vmax and Km. Specifically, Vmax values for free and immobilized laccase were found to be 0.021 mg/L min and 0.045 mg/L min, respectively, while Km values were 147.2 mg/L for free laccase and 136.46 mg/L for immobilized laccase. The lower Km and higher Vmax for immobilized laccase indicate its enhanced affinity towards the substrate, likely due to ultrasound-induced alterations in the enzyme's confirmation and increased exposure of active sites, leading to more efficient degradation. Furthermore, the toxicity and Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that after the treatment process, the wastewater exhibited 70% less toxicity than before treatment, with over 25 compounds degrading by more than 75%. At last, the prepared immobilized laccase had excellent recyclability retaining 70% activity up to 6 consecutive cycles. A straightforward manufacturing strategy and outstanding performance make the recyclable magnetic immobilized Laccase (Fe₃O₄ Laccase) an up-and-coming option for various environmental applications, particularly in water pollution control and treatment.

Keywords: kinetic, laccase enzyme, sonoenzymatic, ultrasound irradiation

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Authors: Kayla Fanning

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Theocritus’ use of near-pastoral motifs in creating the lamenting narrative in “The Sorceress” idyll suggests a link between Simaetha and the quintessential shepherd that ultimately transcends the bucolic serenading structure, evident in “The Serenade”, as it pertains to depictions of women. In Theocritus’ “The Serenade”, an anonymous goatherd serenades his beloved, Amaryllis, in hopes of persuading her to reciprocate his love. This serenade soon turns into a vicious lament where all hope for reciprocation dissolves, leaving the goatherd severely melancholic and malignant. In “The Cyclops’ Serenade”, the cyclops, Polyphemus, sings of Galatea in solitude; in so doing, he negotiates between feelings of heartache and anger that eventually subside. His depiction of Galatea, in being less vindictive than the goatherd’s, manifests less toxicity. In adopting, and essentially creating, this serenading structure, Theocritus illustrates his ability to alter portrayals of women while maintaining the premise of the pastoral serenade; that is the shepherd's lament of his indifferent beloved. A thematic intertextual analysis of the idylls reveals a variety of ways in which the toxicity of the goatherd’s relation to Amaryllis is mutated or even inverted. In “The Sorceress”, a powerful witch named Simaetha spellbinds her unfaithful lover and angrily laments his betrayal in a way that is reminiscent of the goatherd's harmful behavior towards Amaryllis.

Keywords: femininity, pastoral, serenade, Theocritus

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Authors: Azad Khalid, Sifa Dogan

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Because of its persistence in conventional treatment plants and broad prevalence in water bodies, the pharmaceutical chemical carbamazepine (CBZ) has been suggested as an anthropogenic marker to evaluate water quality. This study provides a thorough examination of the origins and occurrences of CBZ in water bodies, as well as the drug's toxicological effects and laws. Given CBZ's well-documented negative consequences on the human body when used medicinally, cautious monitoring in water is advised. CBZ residues in drinking water may enter embryos and newborns via intrauterine exposure or breast-feeding, causing congenital abnormalities and/or neurodevelopmental issues over time. The insufficiency of solo solutions was shown after an in-depth technical study of traditional and sophisticated treatment technologies. Nanofiltration and reverse osmosis membranes are more successful at removing CBZ than traditional activated sludge and membrane bioreactor techniques. Recent research has shown that severe chemical cleaning, which is essential to prevent membrane fouling, may lower long-term removal efficiency. Furthermore, despite the efficacy of activated carbon adsorption and advanced oxidation processes, a few issues such as chemical cost and activated carbon renewal must be carefully examined. Individual technology constraints lead to the benefits of combined and hybrid systems, namely the heterogeneous advanced oxidation process.

Keywords: carbamazepine, occurrence, toxicity, conventical treatment, advanced oxidation process (AOPs)

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Authors: Adewale Tolulope Irewale, Elias Emeka Elemike, Christian O. Dimkpa, Emeka Emmanuel Oguzie

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As the global population steadily approaches the 10billion mark, the world is currently faced with two major challenges among others – accessing sustainable and clean energy, and food security. Accessing cleaner and sustainable energy sources to drive global economy and technological advancement, and feeding the teeming human population require sustainable, innovative, and smart solutions. To solve the food production problem, producers have relied on fertilizers as a way of improving crop productivity. Commercial inorganic fertilizers, which is employed to boost agricultural food production, however, pose significant ecological sustainability and economic problems including soil and water pollution, reduced input efficiency, development of highly resistant weeds, micronutrient deficiency, soil degradation, and increased soil toxicity. These ecological and sustainability concerns have raised uncertainties about the continued effectiveness of conventional fertilizers. With the application of nanotechnology, plant biomass upcycling offers several advantages in greener energy production and sustainable agriculture through reduction of environmental pollution, increasing soil microbial activity, recycling carbon thereby reducing GHG emission, and so forth. This innovative technology has the potential for a circular economy and creating a sustainable agricultural practice. Nanomaterials have the potential to greatly enhance the quality and nutrient composition of organic biomass which in turn, allows for the conversion of biomass into nanofertilizers that are potentially more efficient. Water hyacinth plant harvested from an inland water at Warri, Delta State Nigeria were air-dried and milled into powder form. The dry biomass were used to prepare biochar at a pre-determined temperature in an oxygen deficient atmosphere. Physicochemical analysis of the resulting biochar was carried out to determine its porosity and general morphology using the Scanning Transmission Electron Microscopy (STEM). The functional groups (-COOH, -OH, -NH2, -CN, -C=O) were assessed using the Fourier Transform InfraRed Spectroscopy (FTIR) while the heavy metals (Cr, Cu, Fe, Pb, Mg, Mn) were analyzed using Inductively Coupled Plasma – Optical Emission Spectrometry (ICP-OES). Impregnation of the biochar with nanonutrients were achieved under varied conditions of pH, temperature, nanonutrient concentrations and resident time to achieve optimum adsorption. Adsorption and desorption studies were carried out on the resulting nanofertilizer to determine kinetics for the potential nutrients’ bio-availability to plants when used as green fertilizers. Water hyacinth (Eichhornia crassipes) which is an aggressively invasive aquatic plant known for its rapid growth and profusion is being examined in this research to harness its biomass as a sustainable feedstock to formulate functionalized nano-biochar fertilizers, offering various benefits including water hyacinth biomass upcycling, improved nutrient delivery to crops and aquatic ecosystem remediation. Altogether, this work aims to create output values in the three dimensions of environmental, economic, and social benefits.

Keywords: biochar-based nanofertilizers, eichhornia crassipes, greener agriculture, sustainable ecosystem, water hyacinth

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Authors: Syeda Hira, Muhammad Gulfraz

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Background: Liver diseases cause serious health issues. Plants contain active compounds that significantly help in the treatment of various diseases. Ficus carica is traditionally used for the treatment of liver diseases. The purpose of the present study was the isolation and identification of active components from F.carica leaves which are responsible for hepatoprotective activity. Methods: The study was designed to identify the most active hepatoprotective sub-fraction from ethyl acetate fraction of Ficus carica by in vitro study and evaluation of its in vivo hepatoprotective effect in animal models. Ethyl acetate fraction was subjected to column, and a total of eight sub-fractions were obtained. In vitro, the hepatoprotective effect of all sub-fractions was determined on HepG2 cell lines. Toxicity was induced by CCl₄ (Carbon tetrachloride), and silymarin was used as a positive control. On the basis of the results, the most active sub-fraction was subjected to LC-MS and FT-IR analysis for the identification of bioactive compounds. In vivo, the hepatoprotective effect was determined in mice. Toxicity was induced by CCl₄; at the end of the experiment, biochemical parameters such as ALT, AST, ALP, bilirubin, and total protein were estimated in serum. Histopathology of liver tissues was also done. Results: Sub-fraction FVI exhibited significant (P<0.05) hepatoprotective activity as compared to other sub-fractions, which was almost similar to the standard drug silymarin. Six known bioactive compounds were identified from this sub-fraction after LC-MS analysis. In vivo, the hepatoprotective activity of sub-fraction FVI was evaluated in CCl₄-induced toxicated mice. Administration of CCl₄ significantly increased level of ALT (Alanine transaminase), AST (Aspartate aminotransferase), ALP (Alkaline phosphatase), and bilirubin and decreased the total protein. Treatment with sub-fraction FVI significantly (p<0.05) reversed the level of these biomarkers toward normal at both doses of 25 mg/kg and 50 mg/kg. Conclusion: Our findings confirmed the hepatoprotective effect of ethyl acetate fraction of F.carica. It could be a good candidate for the development of a natural hepatoprotective drug; pre-clinical investigation on ethyl acetate fraction is recommended.

Keywords: Ficus carica, hepatoprotective, CCl₄, bioactive compounds, liver markers

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Authors: Azade Ghorbani-HasanSaraei, Seyed-Ahmad Shahidi, Sakineh Alizadeh, Adeleh Maghsoudlou

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Improvement of color, as a quality attribute of Plum Concentrate, has been made possible by the increase in knowledge of kinetic of color change. Three different heating/evaporation processes were employed for the production of pPlum juice concentrate. The Plum juice was concentrated to a final 55 °Bx from an initial °Bx of 15 by microwave heating, rotary vacuum evaporator and evaporating at atmospheric pressure. The final Plum juice concentration of 55 °Bx was achieved in 17, 24 and 57 min by using the microwave, rotary vacuum and atmospheric heating processes, respectively. The colour change during concentration processes was investigated. Total colour differences, Hunter L, a and b parameters were used to estimate the extent of colour loss. All Hunter colour parameters decreased with time. The zero-order, first-order and a combined kinetics model were applied to the changes in colour parameters. Results indicated that variation in TCD followed both first-order and combined kinetics models, and parameters L, a and b followed only combined model. This model implied that the colour formation and pigment destruction occurred during concentration processes of plum juice.

Keywords: colour, kinetics, concentration, plum juice

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Authors: Tomas Makaras

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Sticklebacks (Gasterosteiformes) are increasingly used in ecological and evolutionary research and become well-established role as model species for biologists. However, ecotoxicology studies concerning behavioural effects in sticklebacks regarding stress responses, mainly induced by chemical mixtures, have hardly been addressed. Moreover, although many authors in their studies emphasised the similarity between three-spined and nine-spined stickleback in morphological, neuroanatomical and behavioural adaptations to environmental changes, several comparative studies have revealed considerable differences between these species in and their susceptibility and resistance to variousstressors in laboratory experiments. The hypothesis of this study was that three-spined and nine-spined stickleback species will demonstrate apparent differences in response patterns and sensitivity to metal-based chemicals stimuli. For this purpose, we investigated the swimming behaviour (including mortality rate based on 96-h LC50 values) of two ecologically similar three-spined (Gasterosteusaculeatus) and nine-spined sticklebacks (Pungitiuspungitius) to short-term (up to 24 h) metal mixture (MIX) exposure. We evaluated the relevance and efficacy of behavioural responses of test species in the early toxicity assessment of chemical mixtures. Fish exposed to six (Zn, Pb, Cd, Cu, Ni and Cr) metals in the mixture were either singled out by the Water Framework Directive as priority or as relevant substances in surface water, which was prepared according to the environmental quality standards (EQSs) of these metals set for inland waters in the European Union (EU) (Directive 2013/39/EU). Based on acute toxicity results, G. aculeatus found to be slightly (1.4-fold) more tolerant of MIX impact than those of P. pungitius specimens. The performed behavioural analysis showed the main effect on the interaction between time, species and treatment variables. Although both species exposed to MIX revealed a decreasing tendency in swimming activity, these species’ responsiveness to MIX was somewhat different. Substantial changes in the activity of G. aculeatus were established after 3-h exposure to MIX solutions, which was 1.43-fold lower, while in the case of P. pungitius, 1.96-fold higher than established 96-h LC50 values for each species. This study demonstrated species-specific differences in response sensitivity to metal-based water pollution, indicating behavioural insensitivity of P. pungitiuscompared to G. aculeatus. While many studies highlight the usefulness and suitability of nine-spined sticklebacks for evolutionary and ecological research, attested by their increasing popularity in these fields, great caution must be exercised when using them as model species in ecotoxicological research to probe metal contamination. Meanwhile, G. aculeatus showed to be a promising bioindicator species in the environmental ecotoxicology field.

Keywords: acute toxicity, comparative behaviour, metal mixture, swimming activity

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Authors: Cunyu Zhang

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Based on Systemic Functional Linguistics, this paper aims to explore the complex aspectuality system of English. This study shows that the complex aspectuality is classified into complex viewpoint aspect which refers to the homogeneous or heterogeneous ways continuously viewing on the same situation by the speaker and complex situation aspect which is the combined configuration of the internal time schemata of situation. Through viewpoint shifting and repeating, the complex viewpoint aspect is formed in two combination ways. Complex situation aspect is combined by the way of hypotactic verbal complex and the limitation of participant and circumstance in a clause.

Keywords: aspect series, complex situation aspect, complex viewpoint aspect, systemic functional linguistics

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Authors: Yohana Fessehazion

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Mercury is a prominent example of a heavy metal contaminant in the environment, and it has been extensively investigated for its potential health risk in humans and other organisms. In South Africa, massive mercury contamination happened in1980s when the England-based mercury reclamation processing plant relocated to Cato Ridge, KwaZulu-Natal Province, and discharged mercury waste into the Mngceweni River. This mercury waste discharge resulted in high mercury concentration that exceeded the acceptable levels in Mngceweni River, Umgeni River, and human hair of the nearby villagers. This environmental issue raised the alarm, and over the years, several environmental assessments were reported the dire environmental crises resulting from the Thor Chemicals (now known as Metallica Chemicals) and urged the immediate removal of the around 3,000 tons of mercury waste stored in the factory storage facility over two decades. Recently theft of some containers with the toxic substance from the Thor Chemicals warehouse and the subsequent fire that ravaged the facility furtherly put the factory on the spot escalating the urgency of left behind deadly mercury waste removal. This project aims to investigate the mercury contamination leaking from an old Thor Chemicals mercury processing plant. The focus will be on sediments, water, terrestrial plants, and aquatic weeds such as the prominent water hyacinth weeds in the nearby water systems of Mngceweni River, Umgeni River, and Inanda Dam as a bio-indicator and phytoremediator for mercury pollution. Samples will be collected in spring around October when the condition is favourable for microbial activity to methylate mercury incorporated in sediments and blooming season for some aquatic weeds, particularly water hyacinth. Samples of soil, sediment, water, terrestrial plant, and aquatic weed will be collected per sample site from the point of source (Thor Chemicals), Mngceweni River, Umgeni River, and the Inanda Dam. One-way analysis of variance (ANOVA) tests will be conducted to determine any significant differences in the Hg concentration among all sampling sites, followed by Least Significant Difference post hoc test to determine if mercury contamination varies with the gradient distance from the source point of pollution. The flow injection atomic spectrometry (FIAS) analysis will also be used to compare the mercury sequestration between the different plant tissues (roots and stems). The principal component analysis is also envisaged for use to determine the relationship between the source of mercury pollution and any of the sampling points (Umgeni and Mngceweni Rivers and the Inanda Dam). All the Hg values will be expressed in µg/L or µg/g in order to compare the result with the previous studies and regulatory standards. Sediments are expected to have relatively higher levels of Hg compared to the soils, and aquatic macrophytes, water hyacinth weeds are expected to accumulate a higher concentration of mercury than terrestrial plants and crops.

Keywords: mercury, phytoremediation, Thor chemicals, water hyacinth

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Authors: Sudhir Kumar, V. R. Sinha

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Background: The topical and systemic toxicity associated with present nonmelanoma skin cancer (NMSC) treatment therapy using 5-Fluorouracil (5-FU) make it necessary to develop a novel delivery system having lesser toxicity and better control over drug release. Solid lipid nanoparticles offer many advantages like: controlled and localized release of entrapped actives, nontoxicity, and better tolerance. Aim:-To investigate safety and efficacy of 5-FU loaded solid lipid nanoparticles as a topical delivery system for the treatment of nonmelanoma skin cancer. Method: Topical solid lipid nanoparticles of 5-FU were prepared using Compritol 888 ATO (Glyceryl behenate) as lipid component and pluronic F68 (Poloxamer 188), Tween 80 (Polysorbate 80), Tyloxapol (4-(1,1,3,3-Tetramethylbutyl) phenol polymer with formaldehyde and oxirane) as surfactants. The SLNs were prepared with emulsification method. Different formulation parameters viz. type and ratio of surfactant, ratio of lipid and ratio of surfactant:lipid were investigated on particle size and drug entrapment efficiency. Results: Characterization of SLNs like–Transmission Electron Microscopy (TEM), Differential Scannig calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), Particle size determination, Polydispersity index, Entrapment efficiency, Drug loading, ex vivo skin permeation and skin retention studies, skin irritation and histopathology studies were performed. TEM results showed that shape of SLNs was spherical with size range 200-500nm. Higher encapsulation efficiency was obtained for batches having higher concentration of surfactant and lipid. It was found maximum 64.3% for SLN-6 batch with size of 400.1±9.22 nm and PDI 0.221±0.031. Optimized SLN batches and marketed 5-FU cream were compared for flux across rat skin and skin drug retention. The lesser flux and higher skin retention was obtained for SLN formulation in comparison to topical 5-FU cream, which ensures less systemic toxicity and better control of drug release across skin. Chronic skin irritation studies lacks serious erythema or inflammation and histopathology studies showed no significant change in physiology of epidermal layers of rat skin. So, these studies suggest that the optimized SLN formulation is efficient then marketed cream and safer for long term NMSC treatment regimens. Conclusion: Topical and systemic toxicity associated with long-term use of 5-FU, in the treatment of NMSC, can be minimized with its controlled release with significant drug retention with minimal flux across skin. The study may provide a better alternate for effective NMSC treatment.

Keywords: 5-FU, topical formulation, solid lipid nanoparticles, non melanoma skin cancer

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Authors: Alyaa Ahmed Farid, Aida Ismail, Ibrahim Rabia, Azza Fahmy, Azza El Amir

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This study was designed for assessment of 3 types of Cysteine protease inhibitors (CPIs) fluromethylketone (FMK), vinyl sulfone (VS) and sodium nitro prussid (SNP), to define which of them is the best? The experiments aimed to define the protective power of each inhibitor alone or combined with PZQ for curing S. mansoni infection in mice. In vitro, treated S. mansoni adult worms recorded a mortality rate after 1 hr of exposure to 500 ppm of FMK, VS and SNP as 75, 70 and 60%, while, treated cercaria recorded 75, 60 and 50%, respectively. FMK+PZQ treatment recorded the maximum reduction in worm burden (97.2% at 5 wk PI). VS treatment alone or combined with PZQ increases IgM, total IgG, IgG2 and IgG4 levels. In EM study of worm tegument, while only detachment of spines was observed in PZQ treated group, the completely implanted spines were reported in the degenerated tegument of adult worms in all groups treated with CPIs. Treatment with VS+PZQ increased Igs levels but, its effect was different on worm reduction. So, it is not enough to eliminate the infection and FMK+PZQ considered the antischistosomicidal drug of choice.

Keywords: praziquantel, fluromethylketone, vinyl sulfone, worm burden, immunoglobulin pattern

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Authors: Caroline Mendes, Mary McNamara, Orla Howe

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Drug delivery systems are proposed for use in cancer treatment to specifically target cancer cells and deliver a therapeutic dose without affecting normal cells. For that purpose, the use of folate receptors (FR) can be considered a key strategy, since they are commonly over-expressed in cancer cells. In this study, cyclodextrins (CD) have being used as vehicles to target FR and deliver the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within their cavities. Here, β-CD has been modified using folic acid so as to specifically target the FR. Thus, this drug delivery system consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 15.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 16.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 10.5 for A549 and 132.6 µM ± 16.1 and 288.1 µM ± 26.3 for BEAS-2B. These results demonstrate that free MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug. The use of cell imaging by confocal microscopy has allowed visualisation of FR targeting in cancer cells, as well as the identification of the interlisation pathway of the drug. Hence, the cellular uptake and internalisation process of this drug delivery system is being addressed.

Keywords: cancer treatment, cyclodextrins, drug delivery, folate receptors, reduced folate carriers

Procedia PDF Downloads 310